Anti-HER2 mAb H2Mab-77 was established as previously described (29 (link)). To generate H77B, we subcloned VH cDNA of H2Mab-77 and CH of Dog IgGB into the pCAG-Ble vector (FUJIFILM Wako Pure Chemical Corporation). VL cDNA of H2Mab-77 and CL cDNA of dog kappa light chain were also subcloned into the pCAG-Neo vector (FUJIFILM Wako Pure Chemical Corporation). The vector of H77B was transduced into BINDS-09 (FUT8-deficient ExpiCHO-S) cells using the ExpiCHO Expression System (Thermo Fisher Scientific, Inc.) (37 (link)–41 (link)). H77Bf was purified using Ab-Capcher (ProteNova Co., Ltd.). Dog IgG was purchased from Jackson ImmunoResearch Laboratories, Inc.
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Generation of Humanized Anti-HER2 Antibody
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Other organizations : Tohoku University, Microbial Chemistry Research Foundation, Yamaguchi University
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Variable analysis
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- Subcloning of V_H cDNA of H2Mab-77 and C_H of dog IgGB into the pCAG-Ble vector
- Subcloning of V_L cDNA of H2Mab-77 and C_L cDNA of dog kappa light chain into the pCAG-Neo vector
- Transduction of the vector of H77B into BINDS-09 (FUT8-deficient ExpiCHO-S) cells
- Production of H77B antibody
- Use of pCAG-Ble and pCAG-Neo vectors
- Use of BINDS-09 (FUT8-deficient ExpiCHO-S) cells
- Purification of H77B using Ab-Capcher
- Dog IgG purchased from Jackson ImmunoResearch Laboratories, Inc.
- Not explicitly mentioned
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